약학회지 제42권 제 1호 12니4(1998)
Yakhak Hoeji Vol. 42, No. 1
3-(치환) 테트라조일메철세파로스포린의 합성과 생리활성
고옥현* ■ 김영수 ■ 고봉석 • 이재영 • 하재천 • 방희재 • 유진철
조선대학교약학대학 (Received October 29. 1997)
강형룡
S y n t h e s i s a n d B i o l o g i c a l A c t i v i t y o f 3 - ( S u b s t i t u t e d ) T e t r a z o l y I m e t h y 1 C e p h a l o s p o r i n s
O k - H y u n Ko*, Y o u n g - S o o K im , B o n g - S u k K o, J a e - Y o u n g Lee, J a i - C h u n H a , H e e - Ja e B a n g . J in - C h e o l Y o o a n d H y u n g - R y o n g K a n g
College o f Pharmacy, Chosun University, Kwangju, 501-759 Korea
Abstract— For the developement of new cephalosporin antibiotics with aminothiazolcarboxymethy- lethoxyimino moiety on the C-7 position and tetrazolymethyl moiety on the C-3 position of cephem ring.
7p-[{Z)-2-(2-aininothiazol-4-yl)-2-(l--carboxy-l-rnethylethoxyimino)acetarnido]-3-[5-(substituted) tetrazol- 2-yl]methyl-3-cephem-4-carboxylic acids(28-35) were synthesized. These compounds were tested for an
timicrobial activity in vitro against Gram(+) and Gram(-) bacteria. They showed remarkable an
tibacterial activity against Escfterichia coli AB 1157. Escherichia coli AB 0111, Escherichia coli BE 1186. Mi
crococcus luteus ATCC 9341. Salmonella typhimurium TV 119, Salmonella typhimurium SL 1102. Staphylococcus au~
rf MS IFO 12732. Staphylococcus aureus R-209. but these compounds were not active against Pseudomonas aeru - ginosa N-10.
Keywords □ Formylation. ACLE. tetrazole. HOBT-DCC method. Vilsmeyer reagent, sillylation. de- formylation, hydrolysis, antibacterial activity.
지금까지의c e p h a l o s p o r i n 계항생제는c e p h e m핵외
C - 7 . C - 3 , C - 4 위치를변형시킨화합물들이다. 0 7위
처에는항균력과G ( - )균의 외막투과성을 증가시켜광
범위 항균s p e c t r u m을갖게하는a m i n o t h i a z o l e기와
p - l a c t a m a s e에안정성을증가시키고p e n i c i l l i n b i n d
i n g p r o t e i n ( P B P )에 대한 결함친화성을 증가시키는
a l k o x y i m i m o기가s y «체로결합된a m i n o t h i a z o l e a l -
k o x y i m i n o m o i e t y을가진화함물돌이 주류를이루고
있다. 특허a l k o x y i m i n o부위에m e t h o x y i m i n o기를가 신 c e f t i x o z i m e 오'크\ c e f o t a x i m e ^ c e f t n e n o x i m e ^ ' 그 러고 c e f t r i a x o n e가등이 보고되었으며 c a r b o x y m e - t h o x y i m i n o 기와c a r b o x y m e t h y l e t h o x y i m i n o 기를각 각가진c e fix im e ® *과c e f t a z i d i m e ^ *돌이우수한광범위
본 논문에 관한 문의는 이 저자에게로 ( 전화) 062-220-3748 (팩스) 062-234-3016
항균력이있다고보고되었다. C - 3위처는항균력. 흡수.
대사를결정짓는중요한부위이다. P s e u c fo m o m z s와같
은G ( - )균에강력한항균력을가지며 p - l a c t a m a s e에
대한 저항성을 향상시킬 목적으로 C - 3위치에 h e t -
e r o c y c l i c - t h i o m e t h y l효산" 비기찬 q u a r t e m a r y a m m o
n i u m v i n y P ■해, c a t e c h o l ^■체들을도입시
킨세파로스포린계항생제둘이보고되어있다. 0 4 위치
Fig. 1 — General structure of aminothiazole-alkoxyimi - nocephalosporin.
12
3-(치환) 테트라조일메칠세파로스포린의 합성과 생러활성 13
의c a r b o x y기에e s t e r룰도입시켜 흡수율을개선하고
생체이용율을높일목적으로p r o d m g형태의c e p h a l o -
s p o r i n계항생제^"젊^도보고되었다.
본저자등은0 7 위처에 a m i n o t h i a z o l e - c a r b o x y -
m e t h y l e t h o x y i m i n o기틀 도입시키고0 3 위처에 약
러활성이 기대되는t e t r a z o l e을도입시킨c e p h a l o s -
p o r i n계항생제를합성하여 G ( + ) 및G ( - )균에대하
여항균력을기존의 항생제와비교실험한결과를보 고하려고한다.
실 험
시약및기기
본 실험에 사용된 시약둘은 A l d r i c h C o . . S i g m a
C o . . T o k y o K a s e i . , F l u k a C o .에서구입한일급시약 을사용하였으며p - m e t h o x y b e n z y l 7 - a m i n ( r 3 - c h l o - r o m e t h y l - 3 - c e p h e m - 4 - c a r b o x y l a t e h y d r o c h l o r i d e ( A C L E )는 O t s u k a사 제품을. ( Z ) - 2 - ( 2 - a m i n o t h i - a z o l ~ 4 - y l ) - 2 - ( 1 - f e r f - b u t o x y c a r b o n y l - 1 - m e t h y l e - t h o x y i m i n o ) a c e t i c a c i d는 L o n z a사 제품을, s i l i c a
g e l ( 2 3 0 - 4 0 0 m e s h )은 S i g m a사 제품을 사용하였고
용매는 필요에 따라 정제하여 사용하였다. M u e l l e r - H i n t o n b r o t h는D i f c o c o . , 제품을사용하였다. T h i n l a y e r c h r o m a t o g r a p h y ( T L C )는K i e s e l g e l F 2 5 4 (0 .2 5
m m )를바른유리판을잘라 이용하였으며t i c s p o t는
자외선램프 U V G L - 5 8을 사용하였다. 융점 측정은
G a l l e n - K a m p 융점측정기를시유하였으며이에대한
보정은하지않았다. C o l u m n c h r o m a t o g r a p h y는s i l - i c a g e l ( 2 3 0 ^ 4 0 0 m e s h , 60A, M e r c k )을사용하였다.
I R s p e c t r a는B r u k e r I F S 6 6을사용하여K B r p e l l e t
으로 측정하였다. N M R s p e c t r a는 t e t r a m e t h y l s i -
l a n e ( T M S )틀내부표준물질로하여B r u k e r F T - 8 0 M H z . F T - 3 0 0 M H z를사용하였다.
(Z)-2-(2-Formamidothiazol-4-yl)-2-(l-ft/t-butoxy- carbonyl-l-methylethoxyiinino)acetic acid(2)
( C H3C0 ) 2 0 1 2 . 4 g ( 0 . 1 2 m o l )과H C O O H 5 . 6 g ( 0 . 1 2
m o l )을 55"^6(rC에서1시간교반하고어름물로냉각하
여 lyC에서 ( Z ) - 2 - ( 2 - a m i n o t h i a z o l - 4 - y l ) - 2 - ( l - t e r f - b u t o x y c a r b o n y l - l - m e t h y l e t h o x y1m i n o ) a c e t i c a c i d
(1) 1 0 . 0 0 g ( 0 . 0 3 m o l )을가하여상온에서 2시간교반하
였다. 반응물에E t O A c 3 0 0 m /을가하여3 0분간교반하
고E t O c증을분리하여감압농측하였다. 잔유물을tso-
p r o p y l e t h e r ( i P E ) 3 0 0 m /에교반하면서분산시켰다. 생
성된 침전을여과하고i P E로수회 세척하였다. P2O5로
건조하여백색결정8 . 9 3 g ( 8 2 % )을얻었다.
I R ( K B r ) c m ' ' ： 3 3 4 7 . 1 7 2 5 , 1 6 3 8 .
' H - N M R ( D M S O r f ^ ) 5 ： 1 . 3 9 ( 9 H . s . C ( C H3)3) . L 4 4 ( 6 H , s , C ( C H3)2) . 7 . 4 K 1 H , s . t h i a z o l e - H ) , 8 .5 2 ( I H . s . H C O ) . 1 2 . 7 6 ( 1 H . b r . s , H C Q N H )
p-Methoxybenzyl 7p-[(Z)-2-(2-formaiiudothiazol-4-yl) -2-(l-/^/f-butoxycarbonyl-l-methyIethoxyimino) aceta- mido] -3-chIoromethyl-3-cephein-4-carboxylate (3)
( M e t h o d A ) 스화합물(2) 3 g ( 8 . 7 4 m m o l ) , 1 - h y - d r o x y b e n z o t r i a z o l e h y d r a t e ( H O B T ) 1 . 3 8 g ( 8 . 9 5 m m o l )을 D M F 2 0 m Z에 용해시키고 N , N - d i c y c l o - h e x y l c a r b o d i i m d e ( D C C ) 1 . 8 5 g (8.9 5 m m o l )을가하 여상온에서 3시간교반후냉장고에 2 4시간방치하였 다. 생성된결정을여과하여 제거하고여액을보관하였 다. 한편A C L E 3 . 5 4 g ( 8 . 7 4 m m o l ) . E t O A c 1 0 0 m / . H2O 2 0 m / . N a H C O a 0 . 7 4 g (8 . 8 m m o l )을 에
O
HCOOH / (CH3C0)20
Scheme I — Synthesis of Compound (2).
Vol 41, No. I 1998
14 고옥현■김영수 - 고봉석 ■이재영■하재천• 방희재• 유진철• 강형룡
서 30분간교반후 EtOAc층을분리하여 20%식염수 20 mZ로세척한후 MgSOi로건조하였다. 이 EtOAc액에 상기보관한반응액을가하여상온에서 3시간교반하였 다. EtOAc층을취하고여액을다시 EtOAc 100 mZ로 추출하여 EtOAc층을 합하여 0.5N-HC1 수용액(50
m l). 5% NaaSzOsCSOmZ), brine의순으로세척하고 MgS04로건조하여감압농측하였다. 잔유물은 n-hex- ane에분산시켜 생성된 침전물을여과하여 백색결정 5.13g을 얻었다. 이결정을 column chromatogra- phy(EtOAc/cyclohexane=l：l)로 정제하여 백색결 정 4.5g(68.7%)을얻었다.
(Method B) ： DMF 2.3 ml와 THF 31.2 m/의흔 합용액에 POC13 3.1 m/(33.6 mmol)룰가하여 -10~
0°C에서 30분 동안교반시킨 Vilsmeyer reagent를 만든 용액에 화합물(2) 10.00 g(30 mmol)을가하여 같은 온도에서 1시간동안 교반하였다. 한편 ACLE 11.34g(28mmol)을 EtOAc 120m/에 현탁시키고 N,0-bis(trimethylsilyl)acetamide(BTSA) 17.3 mZ(70 mmol)틀가하여 용해시킨 용액을 -20°C에서
HCNI t t
HCNi
I NaHCOj
위의용액에가하여같은온도에서 1.5시간동안교반하 였다. 이용액에 EtOAc와 H20(1：2)의혼합용매 300 m/률가하고교반시킨후유기층을분리하여포화중 조액 20 m l. brine 20 m；순으로 세척하고 무수 MgS04로건조하였다. 이를감압농축하여이잔유물 을 n-hexane에분산시킨후생성된결정을여과하였 다. 이것을 col- um chromatography(EtOAc/cy- clohexane=l：l)로 정제하여 백색결정 13.8 g(67.
2%)을얻었다.
IR (KBr) cm ' ： 1788, 1723, 1685
'H-NMR (DMSCHiJ 5： 1.38(9H, s, C(CH3)3).
1.45(6H, s, CKCHsU 3.52(2H, d. CrSH). 3.85(3H, s. OCH3), 4.54(2H, d, CH2CI), 5,19(1H, d, Ce-H), 5.25(2H, s, OCH2), 5.83(1H, dd, CrH), 6.50(1H, s, thiazole-H), 6.84~7.32(4H, m, ArH), 8.8K1H, s, HCO), 9.65(1H. d, CONH). 12.65(1H, br.s, HCONH)
H -
{CH3>3SiHN'
：n ；X거
COOPM6
느
"
COOPMB
Scheme II — Synthesis of Compound 3. Method A. Scheme III — Synthesis of Compound 3, Method B.
J. Pharm. Soc. Korea
3-(처환) 테트라조일메칠세파로스포린의 합성과 생러활성 15
5-(Thiophen-2-yl)-2H-tetrazole(4)
2-Thiophenecarbonitrile 10g(91.6mmol), sodi
um azide 6.58 gdOOmmol). NH4CI 5.44g(100 mmol)을증류한 DMF 90 m/에가하고 13(H140°C 에서 17시간동안가열반응시킨반응물을빙수중에 저으면서가하고 2N-HC1 로 pH 2.0가되도록산성화 시켰다. 생성된결정을여과하고얼옴물로세척한후. 95%-EtOH로 재결정하여 백색 결정 10.25 g(73.53
%)을얻었다. mp ： 197-200°C
오H-NMR (DMSO-de) 5： 7.10(1H, m, thiophene^
H). 7.83-7.95(2H. m. thiophen^H)
5-Diphenylmethyl-2H-tetrazole(5)
Diphenylacetonitrile 3.00 g(15.2 mmol). sodi
um azide 1.1 g(16.7mmol). NH4CI 0.91 g(16.7 mmol)을중류한 DMF 15 m/에가하고(4)와같은방 범으로합성하였다.
수득율： 49.9%
mp ： 170-174°C
으H-NMR (DMSO-rf^) 5；5.95(1H. s. CH). 7.01- 7.54(10H. m. ArH)
5-(4-Chlorophenyl)-2H-tetrazoIe(6)
p-Chlorobenzonitrile 3.00 g(21.6 mmol), so
dium azide 1.56 g(24mmol), NH4CI 1.3 g(24 mmol). 증류한 DMF 20 m/을(4)와같은방법으로합 성하였다.
수득율： 60.9%
mp ： 274-275"C
'H-NMR (DMSCny 5 ： 7.64~7.74(2H, m, ArH), 8.02-8.13(2H, m, ArH)
5-(4-Methylthiophenyl)-2H-tetrazole(7)
4-(Methylthio)benzonitrile 5.10g(34mmol), so
dium azide 2.45g(37.4mmol), NH4CI 2.03g(37.4 mmol). 증류한 DMF 30m/을(4)와같은방범으로합 성하였다.
수득율 : 94.4%
mp ；220-223V
*H-NMR (DMSO-ds) 6： 2.56(3H, s, CH3). 7.42
~7.53(2H, m, ArH), 7.94~8.05(2H, m, ArH)
5-(4-Hydroxyphenyl)-2H-tetrazole(8)
4-Cyanophenol 10.00g(84mmol), sodium azi
de 5.17g(92 mmol). NH4CI 4.49g(92 mmol). 증류 한 DMF 60 m/을(4) 와같온방법으로함성하였다.
수득율： 62%
mp '■ 185°C
'H-NMR (DMSCny 6： 6.97-구08(2H. m. ArH).
7.89--8.00(2H. m. ArH)
5-(2-Methoxybenzyl)-2H-tetrazole(9)
2-Methoxybenzylcyanide 12.20g(83mmol), so
dium azide 5.17 g(92 mmol), NH4CI 4.49g(92 mmol), 증류한 DMF 60 m/을(4)와같은방법으로함 성하였다.
수득율： 61.9%
mp ： 175~178°C
'H-NMR (DMSOA) 5 ： 3.74(2H, (3H, s. OCH3), 7.37(4H, m, ArH)
CH2), 3.78
5-(l,5-Dimethyl-2-pyrrole)-2H-tetrazole(10)
1,5-Dimethyl-2-pyrrolecarbonitrile 5.00 g (42 mmol), sodium azide 3,02 g(46 mmol), NH4CI 2.5 g(46mmol), 증류한 DMF 40 m；을 (4)와같은방법 으로합성하였다.
수율： 50.8%
mp ： 198°C
'H-NMR (DMSO-rfs) 5 ： 2.27(3H, s, ArCHa), 3.89(3H, s, N -C H 3), 6.00~6.05(1H, m, ArH), 6.72~6.77(1H, m, ArH)
5-NaphthyI-2H-tetrazole(l 1)
1-Cyanonaphthalene 10.00 g(65.2 mmol), sodi
um azide 4.66g(71.7mmol). NH4CI 3.83g(71.7 mmol). 중류한 DMF 60 m；을화합물(4)와같은방법 으로합성하였다.
수득율：82.4%
mp ： 167--169"C
'H-NMR (DMSO-d,) 6： 7.58~8.69(7H. m, ArH)
p-Methoxybenzyl 7p-[(Z)-2-(2-formamidothiazol-4-yl) -2-(l-^e/t-butoxycarbonyl-l-methylethoxyimino)aceta-
Vol. 42, 1998
16 고옥현■김영수■고봉석■이재영■하재천■방희재■유진철■강형룡
mido]-3-[5-(thiophen-2-yl)tetrazol-2-yl]inethyl-3-cep- hem-4-carboxylate(12)
M e s C O 10 m/에화합물( 3 ) 1.00 g(l.2 7 mmol)을 녹인후 Nal 0.23 g(l.52 mmol)을가하여 상온에서 1.5시간동안반응시킨용액에화합물(4) 0.21 g(1.40 mmol)과 K2CO3 0.21 g(l.53 mmol)을 넣어 45~
50°C에서 5시간동안교반시켰다. 반응물을여과하고 여액을 감압농축하여 잔사를소량외 Me2C0에용해
시키고이것을H 2 0 / E t O H ( 2：l )의흔함용매 120m/
에분산하여 10분간교반하고 d-HCl로 pH 2.0로산 성화하여 생성된결정을여과하여황색걸정 0.75 담을 얻었다. 이결정을 silica gel(230~400mesh)을사 용하여 column chromatography (CH3CN/T0- luene=l：5)로 정제하여 백색 결정 0.43g(42.3%) 을얻었다.
IR (KBr) cm * ： 1787, 1733, 1615.
'H-NMR (DMSO-ds) 8 ： 1,38(9H, s, CKCH：^ , 1.43(6H, s, C(0 0 3)2). 3.75(3H, s, OCH3), 5.04- 5.27(6H, m, C2-2H, C3-CH2, OCH2), 5.41-5.80 (2H. m, Ce-H, CrH), 6.88~7.82(8H, m, thiazole- H, ArH), 8.49(1H, s, HCO), 9.60(1H, d, CONH), 12.68(1H, br.s, HCONH)
/?-Methoxybenzyl 7p-[(Z)-2-(2-formamidothiazol-4-yl) -2-(l-/e/t-butoxycarbonyl-l-methylethoxyiinino)aceta- mido]-3-(5-diphenylmethyltetrazol-2-yl)methyl-3-cep- hem-4-carboxylate(13)
화합물13-19은화합물12와같은방범으로합성하였다.
수득율： 27.2%
IR (KBr) cm * ： 1787, 1733, 1610.
'H-NMR (DMSO-d,) 5 ： 1.38(9H, s, C(CH3)3).
I.44(6H, s, C(CH3)2), 3.75(3H, s. OCH3), 4.90- 5.25(6H, m, C2-2H, C3-CH2. OCH2), 5.38-5.65 (2H, m, Ce-H, CrH), 6.87~7.40(15H, m, thia- zole-H, ArH), 8.49(1H, s, HCO), 9.62(1H, d, CONH), 12.67(1H, br.s, HCONH)
p-Methoxybenzyl 7p-[(Z)-2-(2-formamidothiazol-4-yl) -2-(l-/eit-butoxycarbonyl-l-methylethoxyiniino)aceta- mido]-3-[5-(4-chlorophenyl)tetrazol-2-yl]methyl-3-cep- hem-4-carboxylate(14)
수득율： 51%
나
COOPM8
Scheme IV — Synthesis of Compounds 12-19.
IR (KBr) cm'* ： 1785, 1730, 1612.
'H-NMR (DMSO-dc) 6 ： 1,38(9H. s. C(CH3)3).
1.44(6H, s, C(CH3)2). 3.74(3H. s, OCH3), 4.98- 5.3K6H, m, C2-2H. C/-CH2. OCH2), 5.47-5.73 (2H, m, Cg-H, C7-H), 6.87~8.03(9H, m. thiazole- H, ArH), 8.49(1H, s, HCO), 9.62(1H, d, CONH).
12.66(1H, br.s, HCONH)
p-Methoxybenzyl 7p-[(Z)-2-(2-formamidothiazol-4-yl) -2-(l-^£/t-butoxycarbonyl-l-methylethoxyimino)aceta- mido]-3-[5-(4-methylthiophenyl)tetrazol-2-yl]methyl-3- cephem-4-carboxylate(15)
수득율； 41.6%
IR (KBr) cm * ： 1787, 1733, 1610.
'H-NMR (DMSO-d,) 6 ： 1.38(9H, s. CKCHjU I.44(6H, s. C(CH3)2). 2.53(3H, s , SCH3), 3.75(3H.
s, OCH3), 5,04~5.46(6H, m, Cr2H, C/HDH2, OC- Hj), 5.53(1H, dd, (VH). 5.66~5.73(1H, m, CrH), 6.88—8.18(9H, m, thiazole-H, ArH), 8.49(1H, s, HCO), 9.62(1H, d, CONH), 12.68(1H, br.s.
HCONH)
Pharm. Soc. Korea
3-(치환) 테트라조일메철세파로스포린의 합성과 생리활성 17
p-Methoxybenzyl 7P- [(Z)-2-(2-formamidothiazol-4-yl) -2-(l-re/t-butoxycarbonyl-l-methylethoxyimino)aceta- mido]-3-[5-(4-hydroxyphenyI)tetrazol-2-yl]methyl-3- cephem-4-carboxylate(16)
수득율： 37%
IR (KBr) cm'* ： 1785, 1733, 1612.
*H-NMR (DMSO-rfs) 8 ： 1.38(9H, s, CKCHaW.
1.45(6H, s, C(CH3)2). 3.76(3H, s, OCH3), 5.99~
5.28(6H, m, C2-2H, C3-CH2. OCH.), 5.52(1H.
dd, Ce-H), 5.58~5.71(1H, m. CtH). 6.89-7.87 (9H, m, thiazole-H, ArH), 8.50(1H, s, HCO), 9.63(1H, d, CONH), 12.69(1H, br.s. HCQNH)
p-Methoxybenzyl 7p-[(Z)-2-(2-formamidothiazol-4-yl) -2-(l-/£/t-butoxycar-bonyl-l-inethylethoxyimino)aceta- inido]-3-[5-(2-methoxybenzyl)tetrazol-2-yl]methyl-3- cephem-4-carboxylate(17)
수득율： 55%
IR (KBr) cm'* ： 1787, 1731, 1612.
'H-NMR (DMSO-dJ 5 ： 1.380H, s, CKCHs)；；), 1.44(6H, s, C(CH3)2). 3.57(2H, s, -CHzAr), 3.76 (6H, s, 2XOCH3), 5.04-5.28(6H, m, C2-2H, C/-CH2, OCH2), 5.47-5.70(2H, m, Ce-H, C7-
H), 6.89-7.86(9H, m, thiazole-H, ArH), 8.49 (IH, s, HCO). 9.62(1H. d, CONH), 12.63(1H, br.s, HCQNH)
p-Methoxybenzyl 7p-[(Z)-2-(2-formamidothiazol-4-yl) -2-(l-^er^butoxycarbonylmethylethoxyimino)aceta- mido]-3-[5-(l,5-diinethyl-2-pyiTole)tetrazol-2-yl]methyl- 3-cephem-4-carboxylate(18)
수득율： 47%
IR (KBr) cm'' ： 1787, 1733, 1594.
'H-NMR (DMSO-rfs) 8 ： 1.38(9H, s, C(CH3)3), 1.44(6H, s, CKCHsU 2.25(3H, s, CHa-Ar), 3.74~
3,76(6H, m, OCH3. N-CH3). 5.01~5,28(6H, m, C2-2H, C3-CH2, OCH2), 5.44(1H, dd. Ce-H), 5.67
~5.70(1H, m, C7-H), 6.64QH, s, thiazole-H), 6.89~7.40(6H, m, ArH), 8.49(1H, s. HCO), 9.61 (IH, d, CONH), 12.68(1H. s, HCQNH)
p-Methoxybenzyl 7P-[(Z)-2-(2-foraiainidothiazol-4-yl)
Vol. 42, No. 1, 1998
-2-(l-/erf-butoxycarbonylmethylethoxyimino)aceta- mido]-3-(5-naphthyltetrazol-2-yl)methyl-3-cephem-4- carboxyIate(19)
수득율： 43%
IR (KBr) cm'' ； 1787, 1733. 1620.
'H-NMR (DMSO-ds) 8 ： 1.370H. s, C(CH3)3).
1.44(6H, s, C(CH3)2). 3.69~3.72(6H. m, OCH3), 5.05~5.29(6H, m, C2-2H, C3-CH2, OCH2), 5.48- 5.72(2H, m, CrH, CtH ), 6.85~8.78(12H, m, thiazole-H, ArH), 8.50(1H, s, HCO) 9.62(1H, d, CONH), 12.69(1H, s, HCQNH)
p-Methoxy benzyl 7p-[(Z)-2-(2-aminothiazol-4-yl)-2- (l-f£rt-butoxycarbonyl-l-methylethoxyimino)acetami- do]-3-[5-(thiophen-2-yl)tetrazol-2-yl]methyl-3-cephem- 4-carboxylate(20)
화함물(12) 0.50g(0.63inmol)에 MeOH 10 m/.
THF 5 m/, c-HCl 0.2 m/을가하여 2.5시간동안상 온에서교반하였다. 5% NaHCOa 용액으로중화시키 고감압농측한후다시 10% HC1 로 pH 2.0으로산성
12 - 19
MeOH / THF / HCl
그
COOPMB
2 0 - 2 7
Scheme V — Synthesis of Compounds 20-27.
18 고옥현• 김영수■고붕석• 이재영■하재천■방희재■유진철■강형룡
화시켰다. EtOAc 100 mZ, H2O 50 m；를가하여유기 층을취하여 brine 100m/(50 m；x2)로세척한후무 수M g S O i로건조하였다. 부^^^농측하여잔유물을 «- hexane에분산하고생성된결정을여과하였다. 황색 결정 0.31 용을얻었다. 이결정을 silica gel(230~400 mesh)을사용하여 column chromatography(CHj- CN/Toluene=l：7)로정제하여백색결정 0.15g(48.
4%)을얻었다.
IR (KBr) cm'* ： 1787, 1733, 1618.
*H-NMR (DMSO-d^ 5 ： 1.39(15H, br.s, CXCftk
C iC W i). 3.75(3H, s. OCH3), 4.99~5.54(6H, m, C2-2H, C/-CH2. OCH2), 5.45~5.72(2H. m, CrH, C7-H), 6.70~7.82(10H, m, thiazole-H, NHz，ArH), 9.60(1H, d, CONH)
p-Methoxybenzyl 7P-[(Z)-2-(2-aniinothiazoI-4-yl)-2- (l-teit-butoxycarbonyi-l-methylethoxyimino)acetaini- do]-3-(5-diphenylmethyItetrazol-2-yl)methyl-3-cep- hem-4-carboxylate(21)
화합물21~27은 화합물20의 방법으로 각각 합성하
였다.
수득율：37%
IR (KBr) cnf* ： 1787. 1731, 1615.
'H-NMR (DMSCH4) 8： 1.37(15H, br.s, CCCH.Ja， C(CH3)2). 3.74(3H, s, OCH3). 4.93~5.25(6H, m,
C2-2H, C/-CH2, OCH2), 5.43~5.64(2H, m, CrH, Cy-H), 6.69~7.33(17H, m, thiazole-H. NH2, ArH), 9.47(1H. d, CONH)
p-Methoxybenzyl 7P-[(Z)-2-(2-aminothiazol-4-yl)-2- (l-^err-butoxycarbonyl-l-methylethoxyimino)acetami- do]-3-[5-(4-chlorophenyl)tetrazol-2-yl]methyl-3-cep- hem-4-carboxylate(22)
수득율： 50%
IR (KBr) cm ' ： 1787, 1735, 1687.
'H-NMR (DMSO-rfs) 5 ： 1,38(15H, br.s. CKCH,)：；, C(CH3)2). 3.74(3H. s , OCH3). 4.98~5.32(6H, m.
C2-2H. C/-CH2. OCH2), 5.47~5.62(1H, m, CrH), 5.73(1H, br.s, CrH). 6.70~8.04(10H, m, thiazole- H, NH2. ArH), 9.46(1H. d, CONH)
p-Methoxybenzyl 7p-[(Z)-2-(2-aminothiazol-4-yl)-2-
(l-rm-butoxycarbony]-l-methylethoxyimino)acetaini- do]-3-[5-(4-methylthiophenyl)tetrazol-2-yl]methyl-3- cephem-4-carfooxylate(23)
수득율： 48.3%
IR (KBr) cm ' ： 1787. 1731, 1615.
'H-NMR (DMSO-d,) 5 ： 1,38(9H, s, CXCHsW, 1.42(6H, s, C(CH3)2). 2.53(3H, s, SCH3), 3.75 (3H, s, OCH3). 4.99~5.33(6H, m. C2-2H, C/- CH2, OCH2), 5.51~5.74(2H. m, CrH, C7-H), 6.71
~7.96(11H, m, thiazole-H, NH2, ArH), 9.47(1H, d, CONH)
p-Methoxybenzyl 7p-[(Z)-2-(2-aminothiazol-4-yl)-2- (l-/£/t-butoxycarbonyl-l-methy]ethoxyimino)acetaini- do]-3-[5-(4-hydroxyphenyI)tetrazol-2-yl]methyl-3-cep- hein-4-carboxylate(24)
수득율： 62.5%
IR (KBr) cm * ： 1785, 1731, 1615.
'H-NMR (DMSO-d,) 6： 1.38(15H, br.s, (XCHsk C(CH3)2), 3.75(3H. s, OCH3), 4.99~5.33(6H, m,
C2-2H, C/-CH2, OCH2), 5.54(1H. dd, Ce-H). 5.68- 5.70(1H, m, C7-H), 6.70~7.86(11H, m, thiazole- H, NH2. ArH), 9.46(1H, d, CONH)
p-Methoxybenzyl 7p-[(Z)-2-(2-aminothiazol-4-yl)-2- (l-rerr-butoxycarbonyl-l-methylethoxyimino)acetaini- do]-3-[5-(2-methoxybenzyl)tetrazol-2-yl]methyl-3-cep- hem-4-carboxylate(25)
수득율： 24%
IR (KBr) cm'* ： 1785, 1730, 1684.
'H-NMR (DMSOd,) 8： 1.38(15H, br.s, C(CH3)3, CCCHg)-,), 3.56(2H, s. CHj-Ar), 3.74(6H, s, OCH3,
A1-OCH3). 5.03~5.32(6H, m, C2-2H, C；-CH2. OCH2). 5.41~5.70(2H, m, Cg-H, CrH), 6.69-7.85 (IIH, m, thiazole-H, NH2. Ai^H), 9.62(1H, d, CONH)
p-Methoxybenzyl 7p-[(Z)-2-(2-aminothiazol-4-yl)-2- (l-fe/t-butoxycarbonyl-l-methylethoxyimino)acetaini- do]-3-[5-(l,5-dimethyl-2-pyrrole)tetrazol-2-yI]methyl-3- cephem-4-carboxylate(26)
수득율：42%
J. Pharm. Soc. Korea
3-(처환) 테트라조일메철세파로스포린의합성과생러활성 19
IR (KBr) cm"' ： 1785. 1733, 1602.
'H-NMR (DMSOds) 5 ： 1.35(15H, brs, C(CH3)3. aC H sU 2.23(3H, s, AiH^Hs). 3.73(3H. s. OCH3), 3.88(3H, s, N-CH3), 5,02~5.29(6H, m, C2-2H, C/- CH2, OCH2), 5.37~5.67(2H, m, CrH, CrH), 6.62
~7,55(9H, m, thiazole-H, NH2. ArH), 9.43(1H, d, CONH)
r t누 :n :；느
C O O H
/^-Methoxybenzyl 7p-[(Z)-2-(2-aminothiazoI-4-yl)-2- (l-fert-butoxycarbonyl-l-methylethoxyiinino)acetami- do]-3-(5-naphthyltetrazol-2-yl)methyl-3-cephem-4-car- boxylate(27)
수득율： 46%
IR (KBr) cm'* ： 1787, 1731, 1618.
'H-NMR (DMSOd,) 5 ； 1,34(9H, s,
1.39(6H, s, C(CH3)2), 3.70(3H, s, OCH3), 4.98- 5.27(6H, m, C2-2H, C/CHz. OCH2), 5.55-5.71 (2H. m, Ce-H, CrH), 6.69~8.76(14H, m, thiazole- H, NH2, ArH), 9.46(1H, d, CONH)
7p-[(Z)-2-(2-Aminothiazol-4-yl)-2-(l-carboxy-l-me- thylethoxyimino)acetainido]-3-[5-(2-thiopheii-2-yl) tetrazol-2-yl]methyl-3-cephem-4-carboxylic acid(28)
화합물(20) 0.50 g(0.64 mmole)에 CH2CI2 2 m/와 anisole 2 m l, TFA 4 m/률 0~5°C에서가하여 1.5시 간동안교반시키고 iPE 50 m；에분산하여생성된결 정을여과하였다. 이결정을 5%-NaHC03 용액에녹이 고 EtOAc 20 mZ로세척한후수층을취해 5°C 이하에 서 10%-HCl로 pH 2.0로산성화하여 생성된결정을 여과하여흑갈색결정 0.34g을얻었다. 이걸정을 sil- ica gel(230~400 mesh)을 사용하여 column chro- matography(CH3CN/H2O = 8： 1) 로정제하고냉동건 조하여황색결정 0.14g(41.2%)을얻었다.
IR (KBr) cm * ： 3341, 1772, 1674, 1636.
'H-NMR (DMSOd,) 8 ： 1.45(6H, s. C(CH3)2).
4.94~5.28(4H, m, Cj-H, C/-H), 5.5K1H, dd, C*；- H), 5.63(1H, m, (VH), 6.76-7.87(6H, m, thia- zole-H, NH2, ArH), 9.63(1H, d, CONH)
7p-[(Z)-2-(2-Aminothiazol-4-yl)-2-(l-carboxy-l-me- thyIethoxyimino)acetamido]-3-(5-diphenylmethyl- tetrazoI-2-yl)methyI-3-cephem-4-carboxylic acid(29)
4, 12 . 20, 28 R = 5 , 13 , 2 1 , 29 R =
6. 14, 22. 30 R = - ^ - c i 7 , 1 5 , 23. 3 1 R =
8, 16. 24, 32 R = - ^ ^ oh 9. 1 7 , 25, 33 R =
10, 18, 26, 34 R = 느 3
Scheme V I— Synthesis of Compounds 28-35.
철
화함물29-35는화합물28의방법에따라합성하였다. 수득율：34%
IR (KBr) cm'' ： 3338, 1772, 1633.
'H-NMR (DMSO-cy 5 ： 1.44(6H, s, C(CH3)2), 4.77-5.15(4H, m, C2-H, C/-H), 5.39(1H, dd, CV H), 5.67(1H, s, Cv-H), 5.95(1H, s, -CHPhz), 6.80 (IH. s, thiazole-H), 6.89~7.45(12H, m, NH2. ArH), 9.62(1H, d, CONH)
7p-[(Z)-2-(2-Aminothiazol-4-yl)-2-(l-carboxy-l-me- thylethoxyimino)acetamido]-3-[5-(4-chlorophenyl) tetrazo!-2-yl]methyl-3-cephem-4-carboxylic acid(30)
수득율： 51.6%
IR (KBr) cm ' ： 3323, 1772, 1635.
'H-NMR (DMS0-4i) 5 ： 1.44(6H, s, € ( ^3)2).
3.99(2H, s. CrH), 5.01-5.32(2H, m, C 3 -H ) , 5.5 (IH, dd, Ce-H ), 5.62-5.8(1H, m, CtH), 6.78 (IH, s, thiazole-H), 7.59~7.61(2H, m, ArH), 7.89
~8.03(2H, m, ArH). 9.62(1H, d, CONH)
7p-[(Z)-2-(2-Aminothiazol-4-yl)-2-(l-carboxy-l-me- Uiylethoxyimino)acetamido]-3-[5-(4-methylthiophenyl) tetrazol-2-yl]methyI-3-cephein-4-carboxylic acid(31)
수득율： 47.8%
Vol. 42, No. 1, 1998
20 고윽현• 김영수■고봉석■이재영■하재천• 방희재• 유진철■강형룡
IR (KBr) cm'* ； 3307, 1777. 1731.
'H-NMR (DMSO-ds) 5 ： 1,49(6H, s, C(CH3)2).
2.54(3H, s, SCHs), 5.03-5.31 (2H, m, C/-H), 5.29~5.93(2H, m, Ce-H, CrH), 6,89~8.00(5H, m, thiazole-H, ArH), 9.69(1H, d, CONH)
7p-[(Z)-2-(2-Aminothiazol-4-yl)-2-(l-carboxy-l-me- thylethoxyimino)acetamido]-3-[5-(4-hydroxyphenyl) tetrazol-2-yl]methyI-3-cephem-4-carboxylic acid(32)
수득율： 32%
IR (KBr) cm ' ： 3307, 1767, 1672.
'H-NMR (DMSO-rf,) 5 ： 1.40(6H, s, CKCHsU 4.98~5.32(2H, m, C；-H), 5.54~5.90(2H, m, Cg-
H, CrH), 6.86~7.89(7H, m, thiazole-H, NHj.
ArH), 9.68(1H, d, CONH)
7p-[(Z)-2-(2-Aminothiazol-4-yl)-2-(l-carboxy-l-ine- thylethoxyimino)acetainido]-3-[5-(2-inethoxybeazyl) tetrazol-2-yl] methyl-3-cephein-4-carboxylic acid(33)
수득율： 43%
IR (KBr) cm-' ： 3330. 1774, 1677.
'H-NMR (DMSO-dJ 8 ： 1.48(6H, s, CCCHsW.
3.56(2H, s, CHrAr), 3,73(3H. s, -O CH 3), 5.02- 5.32(4H. m, CrCHa. C3-CH2), 5.50(1H, dd, C,-
H). 5.69(1H, s, C7-H), 6.78(1H, s, thiazole-H), 7,18~7.42(4H, m, ArH, NH2), 7.81~7.89(2H, m.
ArH). 9.62(1H. d, CONH)
7p-[(Z)-2-(2-Aminothiazol-4-yl)-2-(l-carboxy-l-me- thylethoxyiinino)acetamido]-3-[5-(l,5-diinethyl-2- pyrrole)tetrazol-2-yl]methyl-3-cephem-4-carboxylic acid(34)
수득율： 30%
IR (KBr) cm'* ： 3305, 1777, 1679.
'H-NMR (DMSOrf,) 5 ： 1.40(6H, s, C(CH3)2).
2.25(3H, s. A1-CH3). 3,74(3H, s, N-CH3). 5.03- 5.29(4H, m. Cr2H. C3-CH2), 5.44(1H, dd, Cg-H), 5.68QH, m, ArH), 6.64(1H. s. thiazole-H), 6.88~ 6.94(1H, m, ArH), 7.29~7.41(3H. m. ArH), 9.62 (IH, d, CONH)
7P-[(Z)-2-(2-Aminothiazol-4-yl)-2-(l-carboxy-l-me-
thylethoxyimmo)acetamido]-3-(5-naphthyltetrazol-2- yl)methyl-3-cephein-4-carboxylic add(35)
수득율：42%
IR (KBr) cm'* ： 3320, 1772, 1728.
'H-NMR (DMSOds) 5 ： 1.40(6H, s. C(CH3)2).
5.01~5.37(4H, m, C2-CH2, C3-CH2), 5.62-5.85 (2H, m, Ce-H. C7-H), 6.85~8.79(10H, m, thia- zole-H, ArH). 9.63(1H, d, CONH)
항 균 력 실 험
시 험 균 주
Alcaligenes faecalis KCTC 1004, Bacillus subtilis
ATCC 6633, Escherichia coli AB 1157, Escherichia coli AB 0111, Escherichia coli BE 1186, Micrococcus luteus ATCC 9341, Mycobacterium phlei IFO 3158.
Pseudomonas aeruginosa N-10, Salmonella typhimu - rium TV 119, Salmonella typhimurium SL 1102, Sta
phylococcus aureus IFO 12732, Staphylococcus aureus
R-209
배 지
시험균주외전배양및검정 plate의제조목적으로 Mueller Hinton broth (DIF Co.) 를사용하였다.
항 균 활 성 측 정 법
시 험 균 의 전 배 앙- /4/cfl%e«es faecalis KCTC 1004
외 11개외균은액체배지에서 37°C, 24시간진탕배양 하여사용하였다.
검 정 plate의 제 조 - 화합물28, 29, 30, 31, 31, 32, 33, 34, 35를소량의 DMSO에각각녹인후증류수를가하 여최종 DMSO의농도가 2%(V/V)가되도록하였다. 각각의시료 Im/률 2단계희석법으로 14차례희석하여 영양한천배지 14m/와섞었을때. 최종배지의화합물
28, 29, 30, 31, 31, 32, 33, 34, 35 및 대조 물질 (cefotaxime, cefazolin) 의농도가각각 40, 20, 10, 5, 2.5, 1.25, 0.63. 0.31, 0.16, 0.08, 0.04, 0.02, 0.01, 0.005 g/m；이되도록제조하였다.
항 균 력 관 정 - 각각의시험균주들을검정 plate에접
종한것을 37±2°C에서 18시간배양후육안으로관찰 하여성장이 억제되는항균제외 최소발육저지농도 (Minimum Inhibitory Concentration, MIC)를조
Pharm. Soc. Korea
